Supporting Information Legends 1
2
Figure S1. Fatty acid composition in leaves of ar21 and wild type. 16:1 includes cis and 3
trans-16:1 fatty acid. Values are expressed as means ± SD (n = 3). 4
Figure S2. Confirmation of the point mutation in ar21 by Sanger sequencing. AR21/AR21, 5
wild type; ar21/AR21, heterozygous line; ar21/ar21, mutant line. 6
Figure S3. Seed fatty acid analysis with the ar21 x Col F2 population. Re-sequencing of the 7
point mutation in ar21 with 107 lines from F2 population revealed that there were 26 wild type 8
(AR21/AR21), 58 heterozygous lines (AR21/ar21) and 23 mutant lines (ar21/ar21) (Table S3). 10 9
lines of each genotype were randomly selected for fatty acid composition analysis in mature 10
seeds. (a) Fatty acid composition presented as average values of 10 lines with three replicates for 11
each line carried out on batches of 20 seeds. (b) 18:3 fatty acid composition from individual lines. 12
(c) 20:1 fatty acid composition from individual lines. AR21/AR21, wild type; ar21/AR21, 13
heterozygous lines; ar21/ar21, mutant lines. Values are expressed as means ± SD (n = 3). 14
Figure S4. Quantitative PCR (qPCR) analysis of accD and ACC1. Quantitative RT-15
PCR (qRT-PCR) was conducted with 13 DAF seeds from ar21, i4g1 and wild type (WT). 16
Relative expression in mutants was presented as fold change against WT. Values are expressed 17
as mean ± SD (n=3). *, p value < 0.05 (by Student’s t-test). 18
Figure S5. Fatty acid composition in mature seeds of wild type and i4g2. (a) PCR 19
confirmation of T-DNA insertion in i4g2 mutant. (b) Fatty acid composition in mature seeds of 20
wild type and i4g2. 16:1 includes cis and trans-16:1 fatty acid. Values are expressed as means ± 21
SD (n = 3). 22
Figure S6. Growth phenotype of wild type, i4g1, i4g2 and i4g1/i4g2 mutants. The top panel 23
showed the seedlings at three-weeks after germination. The middle panel showed the 24
representative primary inflorescences. The bottom panel showed the representative siliques. 25
Scale bar, 1cm. 26
Figure S7. Fatty acid composition in seeds and leaves of wild type and mutants. (a) Fatty 27
acid composition in mature seeds of wild type (WT) and i4g1/i4g2 double mutants. (b) Fatty acid 28
composition in three-week old leaves of wild type (WT), ar21, i4g1, i4g2 and i4g1/i4g2. 16:1 29
includes cis and trans-16:1 fatty acid. Values are expressed as means ± SD (n = 3). *, p value < 30
0.05; **, p <0.01; ***, p < 0.001 (by Student’s t-test). 31
Table S1. Summary of genome sequencing data from ar21. SNV, single nucleotide variant. 32
Table S2. Summary of premature stop mutations detected in ar21. SNV, single nucleotide 33
variant. 34
Table S3. Genotyping with the ar21 x Col F2 population. Genomic DNA was extracted from 35
107 lines individually. PCR products were sequenced to identify the point mutation in 36
At5g57870. T, mutation; C, wild type; H, heterzygous. 37
Table S4. Primer pairs used in this study. 38
Data S1. Differentially expressed genes in the developing seeds of ar21 and i4g1. 39
Data S2. Plastidic gene expression in the developing seeds of ar21 and i4g1. 40
Data S3. Differentially expressed ribosomal genes in the developing seeds of ar21 and i4g1. 41
Ribosomal genes were complied according to Sormani et al., 2011. 42
Data S4. Differentially expressed genes pertinent to lipid metabolism in the developing 43
seeds of ar21 and i4g1. 44
Data S5. Lipidomics analysis of developing seeds from ar21 and wild type. Values are means 45
of five replicates. Statistically significant differences (two-tailed Student t-test) were calculated 46
between ar21 and wild type. SD, standard deviation. 47
